Methods for increasing mental well-being and / or happiness

Abstract

The invention provides a method of increasing mental well-being and / or happiness in a subject, the method comprising administering to the subject an effective amount of the bacteria or a component thereof. The invention also provides the use of the bacteria or a component thereof in the manufacture of a medicament for increasing mental well-being and / or happiness in a subject, and a composition comprising the bacteria or a component thereof for use in increasing mental well-being and / or happiness in a subject.

Description

[Technical Field]

[0001] The invention provides a method of increasing mental well-being and / or happiness in a subject, the method comprising administering to the subject an effective amount of the bacteria or a component thereof. The invention also provides the use of the bacteria or a component thereof in the manufacture of a medicament for increasing mental well-being and / or happiness in a subject, and a composition comprising the bacteria or a component thereof for use in increasing mental well-being and / or happiness in a subject. [Background technology]

[0002] For many years, the prevalence of mental health problems and disability-adjusted life years (DALYs) attributable to mental health have been increasing (GBD Mental Disorders Collaborators, 2022). The COVID-19 pandemic may have exacerbated the problem, with more people beginning to suffer from mental health issues (Chatterjee et al., 2020) and reduced well-being (Rossouw et al., 2021).

[0003] It is becoming increasingly clear that dietary habits can affect brain health and mental well-being. However, the exact nature of these relationships remains unclear. For example, probiotics are beneficial microorganisms that confer health benefits when consumed in sufficient amounts (FAO / WHO, 2001). Several in vitro and in vivo studies have been conducted to determine the effects of probiotics on cognitive function in model systems and humans. Some of these studies suggest that the consumption of probiotic preparations can improve cognitive function, stress management, and decision-making (Sivamaruthi et al., 2019). To date, there has been little research on the effects of probiotics on well-being.

[0004] Given the increasing prevalence of mental health problems and decreased mental well-being in human populations (and their associated companion animals), there is a need for agents that promote mental well-being in subjects. Furthermore, given the decline in happiness in the population, there is a need for agents that promote happiness. Summary of the Invention

[0005] The present inventors have developed methods, uses and compositions comprising Lacticaseibacillus rhamnosus strain HN001 or derivatives or components thereof to increase mental well-being in a subject.

[0006] The inventors have developed methods, uses and compositions comprising Lacticaseibacillus rhamnosus strain HN001 or derivatives or components thereof to increase well-being in a subject.

[0007] In one aspect, the present invention provides a method of increasing mental well-being in a subject, the method comprising administering to the subject an effective amount of Lacticaseibacillus rhamnosus HN001, or a derivative or component thereof.

[0008] In one aspect, the present invention provides a method of increasing well-being in a subject, the method comprising administering to the subject an effective amount of Lacticaseibacillus rhamnosus HN001, or a derivative or component thereof.

[0009] In one aspect, the present invention provides a method of treating unhappiness in a subject, the method comprising administering to the subject an effective amount of Lacticaseibacillus rhamnosus HN001, or a derivative or component thereof.

[0010] In one aspect, the present invention provides the use of Lacticaseibacillus rhamnosus HN001, or a derivative or component thereof, in the manufacture of a medicament for increasing mental well-being in a subject.

[0011] In one aspect, the present invention provides the use of Lacticaseibacillus rhamnosus HN001, or a derivative or component thereof, in the manufacture of a medicament for increasing well-being in a subject.

[0012] In one aspect, the present invention provides the use of Lacticaseibacillus rhamnosus HN001, or a derivative or component thereof, in the manufacture of a medicament for treating unhappiness in a subject.

[0013] In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof for use in increasing mental well-being in a subject.

[0014] In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof for use in increasing a sense of well-being in a subject.

[0015] In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof for use in treating unhappiness in a subject.

[0016] In one aspect, the invention provides a method for increasing mental well-being in a subject, the method comprising administering to the subject an effective amount of Lacticaseibacillus rhamnosus or a component thereof comprising the AscI restriction enzyme digestion profile of HN001 shown in Figure 5.

[0017] In one aspect, the invention provides a method for increasing well-being in a subject, the method comprising administering to the subject an effective amount of Lacticaseibacillus rhamnosus or a component thereof comprising the AscI restriction enzyme digestion profile of HN001 shown in Figure 5.

[0018] In one aspect, the present invention provides the use of Lacticaseibacillus rhamnosus, or a component thereof, comprising the AscI restriction enzyme digestion profile of HN001 shown in Figure 5, in the manufacture of a medicament for increasing mental well-being in a subject.

[0019] In one aspect, the present invention provides the use of Lacticaseibacillus rhamnosus, or a component thereof, comprising the AscI restriction enzyme digestion profile of HN001 shown in Figure 5, in the manufacture of a medicament for increasing well-being in a subject.

[0020] In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus or a component thereof comprising the AscI restriction enzyme digestion profile of HN001 shown in Figure 5 for use in increasing mental well-being in a subject.

[0021] In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus, or a component thereof, comprising the AscI restriction enzyme digestion profile of HN001 shown in Figure 5, for use in increasing well-being in a subject.

[0022] Any embodiment herein is to be taken mutatis mutandis to any other embodiment unless otherwise indicated. For example, those skilled in the art will understand that the prebiotic examples outlined for the methods of the invention apply equally to the uses and compositions of the invention.

[0023] The present invention is not to be limited in scope by the specific embodiments described herein, which are intended for the purpose of illustration only. Functionally equivalent products, compositions, and methods are clearly within the scope of the invention described herein.

[0024] Throughout this specification, unless expressly stated otherwise or the context requires otherwise, references to a single step, composition of matter, group of steps or group of compositions of matter should be interpreted as encompassing one and more than one (i.e., one or more) of that step, composition of matter, group of steps or group of compositions of matter.

[0025] The invention will now be described by way of the following non-limiting examples and with reference to the accompanying drawings. [Brief explanation of the drawings]

[0026] [Figure 1] Initial quality control and probiotic viability are shown. Panels show viability before acid treatment, 15 minutes after acid treatment, and 30 minutes after acid treatment. [Figure 2] Initial quality control and probiotic viability are shown. Panels show viability before acid treatment, 15 minutes after acid treatment, and 30 minutes after acid treatment. [Figure 3] The test flow chart is shown below. [Figure 4] Estimated marginal means using repeated measures ANOVA are shown. [Figure 5] Comparison of L. rhamnosus strains by PFGE of restriction endonuclease-AscI-digested genomic DNA: HN001 (lane 1), strain A (lane 2), Lactococcus lactis subsp. lactis strain ILI403 DNA digested with SmaI (lane 3), ATCC 53103 (lane 4), Lcr35 (lane 5), LGG (lane 6), ATCC 7469 (lane 7), and ATCC 7469 (lane 8). Lane numbering is from left (1) to right (8). DETAILED DESCRIPTION OF THE INVENTION

[0027] General Techniques and Definitions Unless expressly defined otherwise, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., prebiotic).

[0028] The term "and / or," e.g., "X and / or Y," is understood to mean either "X and Y" or "X or Y," and is to be interpreted as providing explicit support for both meanings or either meaning.

[0029] Throughout this specification the word "comprise" or variations such as "comprises" or "comprising" should be understood to mean the inclusion of a stated element, integer, or step, or group of elements, integers, or steps, but not the exclusion of any other element, integer, or step, or group of elements, integers, or steps.

[0030] As used herein, unless stated to the contrary, the term "about" refers to + / -10%, more preferably + / -5%, and even more preferably + / -1% of the specified value.

[0031] As used herein, the term "subject" refers to any animal. In one example, the animal is a vertebrate. For example, the animal is a mammal, a bird, a chordate, an amphibian, or a reptile. In some embodiments, the subject is a mammal. In preferred embodiments, the mammal is a human, a human companion animal, or a human-related livestock animal. In some embodiments, the subject is a model animal such as a rat, a mouse, or a guinea pig. In some embodiments, the mammal can be a companion animal such as a cat, a dog, or a horse. In some embodiments, the mammal can be a livestock animal such as a pig, a cow, a horse, a goat, a sheep, and a deer.

[0032] In some embodiments, the subject is a human. In some embodiments, the subject is selected from an infant, a child, an adolescent, an adult, and an elderly person. As used herein, "elderly" refers to an adult aged 65 or older. In some embodiments, the subject is an infant. In some embodiments, the subject is a child. In some embodiments, the subject is an adolescent. In some embodiments, the subject is an adult. In some embodiments, the subject is an elderly person. In some embodiments, the subject is characterized as unhappy or somewhat unhappy using the Oxford Happiness Inventory (OHQ). In some embodiments, the subject is characterized as neutral, moderately happy, or rather happy using the OHQ. In some embodiments, the subject has not been diagnosed with any one or more of anxiety, depression, stress, postpartum depression, and postpartum anxiety. In some embodiments, the subject has not been treated for or recommended for one or more of anxiety, depression, stress, enhancing calm, occasional anxiety, immune suppression or dysfunction, vaginal health, prenatal depression, postpartum depression, and postpartum anxiety. In some embodiments, the subject has not been treated for or recommended for anxiety. In some embodiments, the subject has not been treated for or recommended for depression. In some embodiments, the subject has not been treated for or recommended for stress. In some embodiments, the subject has not been treated for or recommended for enhanced calm. In some embodiments, the subject has not been treated for or recommended for occasional anxiety. In some embodiments, the subject has not been treated for or recommended for immune suppression or dysfunction. In some embodiments, the subject has not been treated for or recommended for vaginal health. In some embodiments, the subject has not been treated for or recommended for prenatal depression. In some embodiments, the subject has not received or been recommended treatment for postpartum depression.In some embodiments, the subject has never received or been recommended treatment for postpartum anxiety.

[0033] In some embodiments, the subject is a female. In some embodiments, the subject is pregnant or postpartum. In some embodiments, the subject is prepartum. In some embodiments, the female is not pregnant. In some embodiments, the female has never been pregnant. In some embodiments, the female is not postpartum. In some embodiments, the subject is not prepartum.

[0034] In some embodiments, the subject is male.

[0035] As used herein, "microorganism" or "microorganisms" refers to microscopic organisms, including bacteria, viruses, fungi, or eukaryotes.

[0036] As used herein, "probiotics" refers to live microorganisms that, when administered in sufficient amounts, confer a health benefit on a subject as described herein.

[0037] As used herein, "psychobiotics" refers to a class of probiotics that produce mental health benefits in subjects (Sharma et al., 2021).

[0038] As used herein, "prebiotic" refers to a substrate selectively utilized by a host microorganism that confers a health benefit.

[0039] As used herein, "postbiotic" refers to a preparation of inactive microorganisms and / or components of microorganisms that directly or indirectly confer a health benefit to a host (e.g., as described in Vinderola et al., 2022). In some embodiments, the inactive microorganisms confer a health benefit to the host directly or indirectly. In some embodiments, the inactive microorganisms confer a health benefit to the host directly. In some embodiments, the inactive microorganisms confer a health benefit to the host indirectly. In some embodiments, one or more components of the microorganisms confer a health benefit to the host. In some embodiments, the postbiotic comprises microorganisms that have been killed (e.g., by heat or chemical treatment).

[0040] As used herein, "component," "microbial component," and "postbiotic component" refer to a cell part (fragment) or a product secreted from a cell. Examples of postbiotic components are described, for example, in Zolkiewicz et al. (2020) and Vinderola et al. (2022). In some embodiments, the component is selected from one or more of cell wall fragments, cell supernatant, cell lysate, intracellular metabolites, secreted metabolites, enzymes, proteins, short-chain fatty acids, vitamins, phenols, amino acids, bacteriocins, extracellular vesicles, and exopolysaccharides. In some embodiments, the component is a cell wall component. In some embodiments, the component is a cell-free supernatant. In some embodiments, the component is a cell lysate. In some embodiments, the component is an intracellular metabolite. In some embodiments, the component is an extracellular metabolite. In some embodiments, the component is an enzyme. In some embodiments, the component is a protein. In some embodiments, the component is a short-chain fatty acid. In some embodiments, the component is a vitamin. In some embodiments, the component is a phenol. In some embodiments, the component is an amino acid. In some embodiments, the component is a bacteriocin. In some embodiments, the component is an extracellular vesicle. In some embodiments, the component is an exopolysaccharide. In embodiments, the secretion product is conditioned medium obtained from culturing bacteria.

[0041] As used herein, the term "derivative" or "derivatives" of Lacticaseibacillus rhamnosus HN001 refers to variants and closely related strains of HN001. In some embodiments, a derivative has the same restriction enzyme profile as HN001 (see, e.g., the AscI restriction enzyme profile shown in Figure 5). In some embodiments, a derivative is plasmid-free (e.g., as described in Zhou et al., 2005).

[0042] As used herein, the term "increase" or "increases" or "increased" or "increasing" refers to having a higher or greater level of a given parameter after application of a method, use, or composition described herein compared to the level of the given parameter at baseline.

[0043] As used herein, the terms "treat," "treating," or "treatment" refer to at least partially achieving a desired therapeutic outcome. In some embodiments, treating includes preventing or delaying the onset of one or more symptoms of unhappiness. In some embodiments, treating includes halting or reducing the onset of one or more symptoms of unhappiness. In some embodiments, treating includes increasing well-being in a subject characterized as "unhappy" or "unhappy," e.g., using the Oxford Well-Being Inventory.

[0044] HN001 As described in PCT Publication No. WO 99 / 10476, Lacticaseibacillus rhamnosus HN001 is available at the Australian Government Analytical Laboratories (AGAL) under deposit number NM97 / 09514 dated August 18, 1997. This Budapest Treaty recognized depository is no longer called AGAL, ​​but is now called the National Measurement Institute of Australia (NMIA), located at 1 / 153 Bertie Street, Port Melbourne, Victoria, Australia 3207.

[0045] HN001 has been used for conditions other than increasing well-being, as described, for example, in WO99 / 10476 and WO2018 / 220429. HN001 has been described, for example, in Slykerman et al. (2017) and Tay et al. (2020). HN001 is commercially available from NZMP as SureStart™, an early childhood nutrition brand, and Nutiani™, a health and wellness solutions brand from Fonterra, a dairy cooperative. HN001 is also sold under the names LactoB HN001™, Nutiani HN001™, and DR20™. In some embodiments, HN001 is administered in a reproductively viable form. In some embodiments, a proportion of HN001 is killed, dissolved, fractionated, or weakened. In some embodiments, HN001 is killed, lysed, compartmentalized, or attenuated.

[0046] Well-being The present invention provides methods, uses, and compositions comprising Lacticaseibacillus rhamnosus HN001 or derivatives or components thereof for increasing well-being in a subject. As used herein, and as defined by the American Psychological Association (APA), "well-being" is a state of happiness and satisfaction, with low levels of distress, generally good physical and mental health and outlook, or a good quality of life. In some embodiments, well-being is mental well-being. As used herein, "mental well-being" refers to an individual's ability to cope with the normal stresses of life and contribute to their community.

[0047] In certain embodiments, the subject's mental well-being is increased by about 5% to about 10%, or about 5% to about 15%, or about 5% to about 20%, or about 5% to about 25%, or about 5% to about 30%, or about 5% to about 35%, or about 5% to about 45%, or about 5% to about 55%, or about 5% to about 65%, or about 5% to about 75%, or about 5% to about 85%, or about 5% to about 95%, or about 100% compared to baseline. In certain embodiments, the subject's mental well-being is increased by about 5% to about 10% compared to baseline. In certain embodiments, the subject's mental well-being is increased by about 5% to about 15% compared to baseline. In certain embodiments, the subject's mental well-being is increased by about 5% to about 20% compared to baseline. In certain embodiments, the subject's mental well-being is increased by about 5% to about 30% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 35% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 45% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 55% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 65% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 75% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 85% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 5% to about 95% compared to baseline. In some embodiments, the subject's mental well-being is increased by about 100% compared to baseline.

[0048] In certain embodiments, mental well-being is increased in a subject by at least 5%, or at least 10%, or at least 15%, or at least 20%, or at least 25%, or at least 30%, or at least 35%, or at least 40%, or at least 45%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100% compared to baseline.

[0049] In some embodiments, mental well-being is increased in a subject by improving cognitive brain function. In some embodiments, mental well-being is increased in a subject by improving a sense of happiness.

[0050] In some embodiments, mental well-being is assessed using the Warwick-Edinburgh Mental Well-Being Scale (WEMWBS) (Tennant et al., 2007). In some embodiments, mental well-being is assessed using the Reduced Warwick-Edinburgh Mental Well-Being Scale (SWEMWBS) (Stewart-Brown et al., 2009).

[0051] feeling of happiness The present invention provides methods, uses, and compositions comprising Lacticaseibacillus rhamnosus HN001 or derivatives or components thereof for increasing happiness in a subject. As used herein, and as defined by the American Psychological Association (APA), "happiness" or "happiness" is a feeling of joy, pleasure, satisfaction, and well-being.

[0052] Those skilled in the art will appreciate that happiness can be measured using any happiness tool known to those skilled in the art, such as the Oxford Happiness Inventory (OHQ) (Argyle et al., 1989), the Subjective Happiness scale (SHS) (Lyubomirsky and Lepper, 1999), the Satisfaction with Life Scale (SLS) (Diener et al., 1985), the Panas Scale (PS) (Watson et al., 1988), the Authentic Happiness Inventory (AHI) (Seligman, 2002), the Psychological Wellbeing Scale (PWS) (Ryff et al., 2007, adapted from Ryff, 1989), and the Fordyce Emotions Questionnaire. It will be appreciated that well-being can be assessed using the Functional Well-Being Questionnaire (FEQ) (Fordyce, 1988). In one embodiment, well-being is assessed using the OHQ. The OHQ is described in more detail in Example 1 herein. The OHQ has a total of 29 items. Each item is scored on a Likert scale from 1 to 6 (1 = very untrue, 2 = very untrue, 3 = somewhat untrue, 4 = somewhat true, 5 = mostly true, 6 = very true). Some items are worded positively and others negatively. Therefore, negative items (1, 3, 12, 13, 16, 18, 21, and 29) should be reverse scored (Hills and Argyle, 2002). The total score is then divided by 29. The interpretation of the final score is as follows: a final score of 1-2 is considered "unhappy," 2-3 is considered "slightly unhappy," 3-4 is considered "neutral," 4 is "slightly or moderately happy," 4-5 is "rather happy," 5-6 is "very happy," and a final score of 6 is considered "too happy" (Hills and Argyle, 2002).

[0053] In some embodiments, well-being is assessed using the SHS. In some embodiments, well-being is assessed using the SLS. In some embodiments, well-being is assessed using the PS. In some embodiments, well-being is assessed using the AHI. In some embodiments, well-being is assessed using the PWS. In some embodiments, well-being is assessed using the FEQ.

[0054] In some embodiments, the subject experiences an increase in sense of well-being of about 5% to about 10%, or about 5% to about 15%, or about 5% to about 20%, or about 5% to about 25%, or about 5% to about 30%, or about 5% to about 35%, or about 5% to about 45%, or about 5% to about 55%, or about 5% to about 65%, or about 5% to about 75%, or about 5% to about 85%, or about 5% to about 95%, or about 100% compared to baseline. In some embodiments, the subject experiences an increase in sense of well-being of about 5% to about 10% compared to baseline. In some embodiments, the subject experiences an increase in sense of well-being of about 5% to about 15% compared to baseline. In some embodiments, the subject experiences an increase in sense of well-being of about 5% to about 20% compared to baseline. In some embodiments, the subject experiences an increase in sense of well-being of about 5% to about 30% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 35% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 45% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 55% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 65% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 75% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 85% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 5% to about 95% compared to baseline. In some embodiments, the subject experiences an increase in happiness of about 100% compared to baseline.

[0055] In certain embodiments, the sense of well-being is increased in the subject by at least 5%, or at least 10%, or at least 15%, or at least 20%, or at least 25%, or at least 30%, or at least 35%, or at least 40%, or at least 45%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100% compared to baseline.

[0056] In some embodiments, increasing well-being comprises increasing a subject's score on the OHQ. For example, after application of a method, use, or composition described herein, a subject with an OHQ baseline score of 1 is increased to a score greater than 1, or a subject with a baseline score of 2 is increased to a score greater than 2, or a subject with a baseline score of 3 is increased to a score greater than 3, or a subject with a baseline score of 4 is increased to a score greater than 4.

[0057] In some embodiments, increasing happiness comprises increasing happiness in a neutral or happy subject. In some embodiments, at baseline, the subject is characterized as neutral, moderately happy, or even happy using the OHQ. In some embodiments, increasing happiness comprises increasing a subject with a baseline OHQ score of 3 to a score greater than 3. In some embodiments, increasing happiness comprises increasing a subject with a baseline OHQ score of 4 to a score greater than 4. In some embodiments, increasing happiness comprises increasing a subject with a baseline OHQ score of 5 to a score greater than 5.

[0058] In some embodiments, increasing happiness comprises treating unhappiness in a subject. In some embodiments, at baseline, the subject is characterized as unhappy or somewhat unhappy using the OHQ. In some embodiments, treating unhappiness comprises increasing a subject with a baseline OHQ score of 1 to a score greater than 1. In some embodiments, treating unhappiness comprises increasing a subject with a baseline OHQ score of 2 to a score greater than 2.

[0059] In some embodiments, increasing the sense of well-being in the subject comprises a change in salivary cortisol levels. In some embodiments, the change in salivary cortisol levels is a decrease in salivary cortisol levels. In some embodiments, increasing the sense of well-being in the subject does not comprise a change in salivary cortisol levels.

[0060] Those skilled in the art will appreciate that happiness is a parameter that can be measured separately from anxiety, depression, and stress. In some embodiments, increasing happiness does not include reducing anxiety. In some embodiments, increasing happiness does not include reducing depression. In some embodiments, increasing happiness does not include reducing stress. In some embodiments, increasing happiness does not include reducing postpartum depression. In some embodiments, increasing happiness does not include reducing postpartum anxiety.

[0061] In some embodiments, the method for increasing well-being additionally comprises dietary modification. In some embodiments, the method for increasing well-being does not comprise dietary modification.

[0062] stress As used herein and as defined by the APA, "stress" is a physiological or psychological response to an internal or external stressor. Stress involves changes that affect nearly every system in the body, affecting how people feel and behave. For example, it can manifest as palpitations, sweating, dry mouth, shortness of breath, irritability, rapid speech, increased negative emotions (if already experienced), and prolonged stress fatigue. Severe stress is manifested by general adaptation syndrome. By causing these mind-body changes, stress directly contributes to psychological and physiological disorders and diseases, affecting mental and physical health and reducing quality of life.

[0063] anxiety As used herein and as defined by the APA, "anxiety" is an emotion characterized by nervous apprehension and physical symptoms in which an individual anticipates imminent danger, catastrophe, or disaster. The body often mobilizes itself to deal with the perceived threat, with muscles tensing, breathing quickening, and the heart beating faster.

[0064] Both stress and anxiety are emotional responses; however, stress is usually caused by an external trigger. The trigger can be short-term, such as a work deadline or a fight with a loved one, or long-term, such as being unable to work, discrimination, or a chronic illness. Anxiety, on the other hand, is defined by persistent, excessive worry that does not go away even in the absence of the stressor.

[0065] depression As used herein and as defined by the APA, "major depressive disorder" or "depression" is a serious mood disorder characterized by persistent feelings of sadness, emptiness, pessimism, and / or downcast mood that negatively impact daily activities, including eating and work. Symptoms include feelings of hopelessness, irritability, guilt, worthlessness, or helplessness. It may also include loss of interest or enjoyment in leisure activities, reduced energy, fatigue, or restlessness, leading to difficulties with concentration, focus, memory, and decision-making. Depression may also disrupt normal appetite and / or weight and may lead to suicide or suicide attempts. In addition, depression may be accompanied by digestive disorders and other digestive problems, as well as other pain or distress without a clear physical cause. Postpartum depression (PND, also known as perinatal depression or postpartum depression) shares some characteristics with major depressive episodes; however, PND is characterized by a set of features that distinguish it from major depressive disorder, in addition to the clear connection between PND and pregnancy / childbirth. For example, PND is associated with hormonal changes specific to the transition from pregnancy to the postpartum period. Animal model experiments have shown that rats given hormones to mimic this transition develop behavioral changes typical of PND. In addition, PND is associated with certain psychosocial stressors, including infant temperament, and the stress associated with caring for a newborn. In contrast to typical patients with major depressive disorder, PND patients show reduced activation in many neural regions, such as a blunted amygdala response to negative stimuli unrelated to the infant, or reduced emotional response and regulation in response to the crying of their own infant.

[0066] Calmness As used herein, "calmness" or "calm" is a mental state of ease, free from agitation, excitement, or disturbance. It also refers to being in a state of serenity, quietness, or peace. Calmness occurs most easily in the average person during a state of relaxation, but it can also be found during states of much greater alertness and awareness.

[0067] Other conditions / conditions Additionally, or alternatively, the methods, uses, and compositions described herein can be used to result in one or more of the following in a subject: increasing cognitive function, improving the subject's mood, and reducing cognitive fatigue.

[0068] Additionally or alternatively, the methods, uses, and compositions described herein increase cognitive function in a subject. As used herein and as defined by the APA, "cognitive function" refers to the ability of the mental processes of perception, learning, memory, understanding, awareness, reasoning, judgment, intuition, and language. Additionally or alternatively, the methods, uses, and compositions described herein improve mood in a subject. Additionally or alternatively, the methods, uses, and compositions described herein reduce cognitive fatigue.

[0069] Compositions and Products In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof for use in increasing mental well-being in a subject. In one aspect, the present invention provides a composition comprising Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof for use in increasing a sense of happiness in a subject. In one embodiment, the composition is a food product or a dietary supplement.

[0070] In certain embodiments, the composition is a pharmaceutical composition.

[0071] In some embodiments, the composition is a probiotic composition. In some embodiments, the probiotic composition is a psychobiotic composition. In some embodiments, the composition is a postbiotic composition. In some embodiments, the composition further comprises a prebiotic. In some embodiments, the composition is a combined prebiotic and probiotic composition. In some embodiments, the composition is a combined prebiotic and postbiotic composition. In some embodiments, the composition is a synbiotic composition.

[0072] In some embodiments, the compositions comprise one or more other probiotic or postbiotic microorganisms. In some embodiments, the compositions described herein are suitable for administration to humans.

[0073] In some embodiments, the composition is in a form selected from a tablet, powder, capsule, liquid, syrup, gummy, and sachet.

[0074] In some embodiments, the composition comprises milk lipids, phospholipids, powdered or fresh skim milk, recombinant or reconstituted whole milk powder or skim milk powder, skim milk concentrate, skim milk retentate, concentrated milk, ultrafiltered milk retentate, milk protein concentrate (MPC), milk protein isolate (MPI), calcium-depleted milk protein concentrate (MPC), reduced fat milk, reduced fat milk protein concentrate (MPC), casein, caseinate, milk fat, cream, butter, ghee, anhydrous milk fat (AMF), buttermilk, butterserum, betaserum, hard milk fat fraction, soft milk fat fraction, sphingolipid fraction, milk fat globule membrane fraction, milk fat globule membrane lipid fraction, phospholipid fraction, complex lipid fraction, milk extracellular vesicle fraction, colostrum, colostrum fraction, colostrum protein concentrate (CPC), colostrum whey, immunoglobulins from colostrum fractions, whey (including sweet whey, lactic acid whey, mineral acid whey, or reconstituted whey powder), whey protein isolate (WPI), whey protein concentrate (WPC including whey protein phospholipid concentrate (WPPC)), glycomacropeptide, lactoferrin, iron-lactoferrin, functional lactoferrin variants, functional lactoferrin fragments, vitamin D, calcium, compositions derived from any milk or colostrum process stream, blueberry, bilberry, black currant, extracellular vesicles, miRNA, polyunsaturated fatty acids, curcumin, green tea, fucoidan, silyosoflavones, conjugated linoleic acid, and compositions derived from the retentate or permeate obtained by ultrafiltration or microfiltration of any milk or colostrum process stream.

[0075] The composition may also include other nutrients such as proteins, carbohydrates, vitamins, minerals, or amino acids, hi certain embodiments, the amino acids are arginine and / or glutamine.

[0076] In some embodiments, the composition comprises one or more of coconut flour, vitamin C, and calcium. In some embodiments, the composition comprises coconut flour. In some embodiments, the composition comprises vitamin C. In some embodiments, the composition comprises calcium.

[0077] The food product can be any edible consumer product that can have Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof. Examples of such consumer products include, but are not limited to, cultured milk, yogurt, cheese, milk drinks and yogurt drinks, or milk powder, baby food, fruit juice, rice pudding, rusks, purees, cereals, porridge, confectionery products, reconstituted fruit products, snack bars, food bars, muesli bars, spreads, sauces, dips, sports supplements, including dairy and non-dairy based sports supplements, food additives such as protein sprinkles, dietary supplements, including daily supplement tablets, weaning foods and yogurt, and infant formula, such as breast milk infant formula, in powder or liquid form, including hypoallergenic embodiments of such compositions.

[0078] In some embodiments, the food product is a dairy product. In some embodiments, the dairy product is selected from milk, yogurt, drinking yogurt, butter, cream, ice cream, and cheese. In some embodiments, the food product is a dairy imitation. In some embodiments, the food or nutritional supplement is selected from infant formula, follow-on formula, growth formula, breast milk formula, breast milk supplement, adult nutritional formula, and geriatric nutritional formula.

[0079] Flavorings, colorants, and other additives, carriers, or excipients may also be included in the compositions of the present invention, as is well known to those skilled in the art. It will be understood that a wide variety of additives or carriers may be included in such compositions or foods, for example, to improve or maintain bacterial viability or to increase the therapeutic efficacy of Lacticaseibacillus rhamnosus HN001 or its derivatives. For example, additives such as surfactants, humectants, water-retaining agents, adhesives, dispersants, stabilizers, and osmotic agents may be included, as well as so-called stress additives (such as potassium chloride, glycerol, sodium chloride, and glucose) to improve the vitality, growth, replication, and viability of bacterial cells, as well as cryoprotectants such as maltodextrin. Additives may also include compositions that aid in maintaining the viability of microorganisms during long-term storage, such as unrefined corn oil or "invert" emulsion compositions containing an outer layer of oil and wax and an inner layer of a mixture of water, sodium alginate, and bacteria.

[0080] The compositions of the present invention may contain live Lacticaseibacillus rhamnosus HN001 or a derivative thereof, preferably in a form and amount that allows for viable reproduction. Methods for producing such compositions are well known in the art. The compositions may also contain a carbohydrate source, such as a disaccharide, including sucrose, fructose, glucose, or dextrose. Preferably, the carbohydrate source is one that can be utilized aerobically or anaerobically by Lacticaseibacillus rhamnosus HN001 or a derivative thereof. The compositions may support the reproductive viability of Lacticaseibacillus rhamnosus HN001 or a derivative thereof for a period of preferably greater than about 2 weeks, preferably greater than about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, more preferably greater than about 6 months, and most preferably at least about 2 to about 3 years.

[0081] Methods for producing the compositions of the present invention are well known in the art and may employ standard microbiological and pharmaceutical practices.

[0082] In one embodiment, the composition comprises about 0.1, 0.2, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, 99.5, 99.8, or 99.9% by weight of Lacticaseibacillus rhamnosus HN001 or a derivative or component thereof.

[0083] It will be apparent that the concentration of Lacticaseibacillus rhamnosus HN001 or a derivative thereof in a composition formulated for administration may be lower than the concentration in a composition formulated for, for example, distribution or storage, and the concentration in the composition formulated for storage, and subsequent formulation into a composition suitable for administration, must be sufficient to enable the composition for administration to be suitably concentrated so that it can be administered in a therapeutically effective dose.

[0084] Other probiotic or postbiotic microorganisms Those skilled in the art will understand that the methods, compositions, and products described herein may include one or more other probiotic or postbiotic microorganisms. In some embodiments, the one or more other probiotic or postbiotic microorganisms have beneficial effects on one or more of the respiratory tract, the digestive tract, and the skin. In some embodiments, the one or more other probiotic or postbiotic microorganisms are yeast or bacteria.

[0085] In certain embodiments, the one or more other probiotic or postbiotic microorganisms are selected from the group consisting of Acetilactobacillus, Agrilactobacillus, Akkermansia, Amylolactobacillus, Anaerobutyricum, Apilactobacillus, Bacillus, Bacteroides, Bombilactobacillus, Christensenella, Clostridium, Companilactobacillus, Dellaglioa, Enterococcus, Eubacterium, Faecalibacterium, Fructilactobacillus, Furfurilactobacillus, Holzapfelia, L acticaseibacillus, Lactiplantibacillus, Lactococcus, Lapidilactobacillus, Latilactobacillus, Lentilactobacillus, Leuconostoc, Levilactobacillus, Ligilactobacillus, Limosilactobacillus llus, Liquorilactobacillus, Loigolactobacillus, Parabacteroides, Paucilactobacillus, Pediococcus, Prevotella, Schleiferilactobacillus, Secundilactobacillus, and Streptococcus.

[0086] In one embodiment, the one or more other probiotic or postbiotic microorganisms are Akkermansia muciniphila, Anaerobutyricum hallii, Bacillus coagulans, Bacillus subtilis, Bacteroides fragilis, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium longum, Christensenella minuta, Clostridium butyricum, Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Faecalibacterium prausnitzii, Lacticaseibacillus casei, Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, Lactiplantibacillus pentosus, Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactococcus lactis, Latilactobacillus sakei, Lentilactobacillus kefiri、Leuconostoc mesenteroides、Levilactobacillus brevis、Ligilactobacillus salivarius、LimosilactobacillusIt is selected from fermentum, Limosilactobacillus reuteri, Parabacteroides goldsteinii, Pediococcus acidilactici, Pediococcus pentosaceus, Prevotella copri, Saccharomyces boulardii, and Streptococcus thermophilus.

[0087] In certain embodiments, one or more other lactic acid bacteria are selected from L. reuteri RC-14, B. longum BB536, L. lactis MG1363, L. plantarum LP299V, L plantarum LP-115, B. lactis HN019, L. reuteri JCM1112, L. casei ATCC393, B. animalis B94, L. paracasei LPC-37, L. acidophilus NCFM, B. lactis BL-04, B. lactis Bi-07, L. crispatus LBV88, L. gasseri LG-36, B. longum CCFM1029, L. rhamnosus HN067, L. rhamnosus GG (LGG), L. acidophilus LA-14, B breve M-16V, B. lactis BB12, and L. acidophilus HN017.

[0088] <"0000337">The specific strains included are L. reuteri RC-14, B. longum BB536, L. lactis MG1363, L. plantarum LP299V, and L. plantarum LP-115, B. lactis HN019, L. reuteri JCM1112, L. casei ATCC393, B. animalis B94, L. paracasei LPC-37, L. acidophilus NCFM, B. lactis BL-04, B. lactis Bi-07, L. crispatus LBV88, L. gasseri LG-36, B. longum CCFM1029, L. rhamnosus HN067, L. rhamnosus GG (LGG), B. breve M-16V, B. lactis BB12, and L. acidophilus HN017 is smooth.

[0089] In some embodiments, the one or more other lactic acid bacteria are L. reuteri RC-14. In some embodiments, the one or more other lactic acid bacteria are B. longum. In some embodiments, the one or more other lactic acid bacteria are BB536. In some embodiments, the one or more other lactic acid bacteria are L. lactis MG1363. In some embodiments, the one or more other lactic acid bacteria are L. plantarum LP299V. In some embodiments, the one or more other lactic acid bacteria are L. plantarum LP-115. In some embodiments, the one or more other lactic acid bacteria are B. lactis HN019. In some embodiments, the one or more other lactic acid bacteria are L. reuteri JCM1112. In some embodiments, the one or more other lactic acid bacteria are L. casei ATCC393. In some embodiments, the one or more other lactic acid bacteria are B. animalis B94. In some embodiments, the one or more other lactic acid bacteria are L. paracasei LPC-37. In some embodiments, the one or more other lactic acid bacteria are L. acidophilus NCFM. In some embodiments, the one or more other lactic acid bacteria are B. lactis BL-04. In some embodiments, the one or more other lactic acid bacteria are B. lactis Bi-07. In some embodiments, the one or more other lactic acid bacteria are L. crispatus LBV88. In some embodiments, the one or more other lactic acid bacteria are L. gasseri LG-36. In some embodiments, the one or more other lactic acid bacteria are B. longum CCFM1029. In some embodiments, the one or more other lactic acid bacteria are L. rhamnosus HN067. In some embodiments, the one or more other lactic acid bacteria are L. rhamnosus GG (LGG). In some embodiments, the one or more other lactic acid bacteria are B. breve M-16V. In some embodiments, the one or more other lactic acid bacteria are B. lactis BB12. In one embodiment, the one or more other lactic acid bacteria is L. acidophilus HN017.

[0090] In one embodiment, the one or more other lactic acid bacteria is not L. acidophilus LA-14.

[0091] In some embodiments, the one or more other probiotics are psychobiotics. Examples of psychobiotics are described, for example, in Sharma et al. (2021).

[0092] In some embodiments, the methods, compositions, and products described herein may include one or more prebiotics. Examples of prebiotics are described, for example, in Davani-Davari et al. (2019). In some embodiments, the prebiotic is selected from one or more of galacto-oligosaccharides, fructo-oligosaccharides, trans-galacto-oligosaccharides, inulin, human milk oligosaccharides, gentio-oligosaccharides, xylo-oligosaccharides, isomalto-oligosaccharides, bran, resistant starch, lactulose, lactitol, flavanols, and pectin oligosaccharides. In some embodiments, the prebiotic is human milk oligosaccharide (e.g., as described in Corona et al., 2021). In some embodiments, the human milk oligosaccharides are selected from one or more of 2'-fucosyllactose (2'FL), (3-fucosyllactose) 3FL, (lacto-N-tetraose) LNT, (lacto-N-neotetraose) LNnT, (6-'sialyllactose) 6'SL, (3'-sialyllactose) 3'SL, (6'-sialyllactosamine) 6'SLN, and difucosyllactose (DFL).

[0093] Administration Various routes of administration are possible for the methods, uses, and compositions described herein, including, but not limited to, oral, dietary, rectal, and topical administration. In some embodiments, administration is oral administration. In some embodiments, administration is rectal administration selected from one or more of suppositories, enemas, via colonoscopy or other medical devices, and fecal transplants. In some embodiments, administration is topical administration.

[0094] In some embodiments, administration is daily. In some embodiments, administration is twice daily. In some embodiments, administration is three times daily. In some embodiments, administration is for about 17 days to about 80 days. In some embodiments, administration is for about 17 days to about 70 days. In some embodiments, administration is for about 20 days to about 65 days. In some embodiments, administration is for about 25 days to about 65 days. In some embodiments, administration is for about 30 days to about 60 days. In some embodiments, administration is for at least about 30 days. In some embodiments, administration is for at least about 60 days.

[0095] In some embodiments, administration is for about 2 weeks to about 10 weeks. In some embodiments, administration is for about 3 weeks to about 9 weeks. In some embodiments, administration is for about 4 weeks to about 8 weeks. In some embodiments, administration is for at least 4 weeks, or 5 weeks, or 6 weeks, or at least 7 weeks, or at least 8 weeks. In some embodiments, administration is for at least 4 weeks. In some embodiments, administration is for at least 5 weeks. In some embodiments, administration is for at least 6 weeks. In some embodiments, administration is for at least 7 weeks. In some embodiments, administration is for at least 8 weeks.

[0096] In one embodiment, administration is about 1 x 10 5 ~Approx. 1×10 12 In one embodiment, administration is in the range of about 1 x 10 colony forming units (CFU) of HN001 or a derivative thereof per day. 5 ~Approx. 1×10 11 In one embodiment, the administration ranges from about 1 x 10 CFU of HN001 or a derivative thereof per day. 5 ~Approx. 1×10 10 In one embodiment, the administration ranges from about 1 x 10 CFU of HN001 or a derivative thereof per day. 6 ~Approx. 1×10 10 In one embodiment, the administration ranges from about 1 x 10 CFU of HN001 or a derivative thereof per day. 7 ~Approx. 1×10 10In one embodiment, the administration ranges from about 1 x 10 CFU of HN001 or a derivative thereof per day. 8 ~Approx. 1×10 10 In one embodiment, administration ranges from about 2 x 10 CFU of HN001 or a derivative thereof per day. 9 ~Approx. 1×10 10 In one embodiment, administration ranges from about 4 x 10 CFU of HN001 or a derivative thereof per day. 9 ~Approx. 1×10 10 In one embodiment, administration ranges from about 5×10 CFU of HN001 or a derivative thereof per day. 9 ~Approx. 1×10 10 In one embodiment, administration ranges from about 6 x 10 CFU of HN001 or a derivative thereof per day. 9 ~Approx. 1×10 10 In one embodiment, administration ranges from about 6 x 10 CFU of HN001 or a derivative thereof per day. 9 ~Approx. 9.5×10 9 In one embodiment, administration ranges from about 6 x 10 CFU of HN001 or a derivative thereof per day. 9 In one embodiment, administration comprises about 9.5 x 10 CFU per day. 9 In one embodiment, administration comprises about 9.5 x 10 CFU per day. 10 Contains CFU.

[0097] In one embodiment, administration is about 2 x 10 per day for about 17 to about 80 days. 9 ~Approx. 1×10 10 In one embodiment, administration ranges from about 3 x 10 CFU of HN001 or a derivative thereof per day for about 17 to about 80 days. 9 ~Approx. 1×10 10 In one embodiment, administration ranges from about 4 x 10 CFU of HN001 or a derivative thereof per day for about 17 to about 80 days. 9 ~Approx. 9.8×10 9 In one embodiment, administration ranges from about 5 x 10 CFU of HN001 or a derivative thereof per day for about 17 to about 80 days. 9 ~Approx. 9.7×10 9In one embodiment, administration ranges from about 6 x 10 CFU of HN001 or a derivative thereof per day for about 17 to about 80 days. 9 ~Approx. 9.5×10 9 In one embodiment, administration ranges from about 6 x 10 CFU of HN001 or a derivative thereof per day for about 17 to about 80 days. 9 In one embodiment, administration comprises about 9 x 10 CFU of HN001 or a derivative thereof per day for about 17 to about 80 days. 9 Contains CFU of HN001 or its derivatives.

[0098] Those skilled in the art will understand that probiotics can be produced at a set CFU, but the viability of the probiotics can decrease over time.In some instances, the viability of probiotics can decrease by up to 40% over a 12-month period.The above dosage ranges define the viable cells that are delivered to the subject.

[0099] In some embodiments, the composition contains about 9×10 9 CFU of HN001 or a derivative thereof per day 9 It is manufactured to ensure delivery of CFU of HN001 or its derivatives.

[0100] When used in combination with another therapeutic agent (e.g., one or more other probiotics, postbiotics, prebiotics, or psychobiotics), administration of the compositions useful herein and the other therapeutic agent can be simultaneous or sequential. Simultaneous administration includes administration of a single dosage form containing all components, or administration of separate dosage forms at substantially the same time. Sequential administration preferably includes administration according to different schedules such that there is an overlap in the period between when the compositions useful herein and the other therapeutic agent are provided. [Example]

[0101] Example 1 - Effect of HN001 on happiness Over the years, the prevalence of mental health problems and disability-adjusted life years (DALYs) attributable to mental health have been increasing. The aim of this study was to investigate the survival of HN001 in acidic medium and to analyze consumer feedback after the ingestion of a specifically designed probiotic intervention to assess the role of HN001 on mental health, specifically their well-being scores over a 60-day period. The probiotic strain L. rhamnosus HN001 was administered at 6 × 10 9 A capsule containing a dose of colony-forming units (CFU) was administered daily. The primary outcome was the change in Oxford Well-Being Questionnaire (OHQ) scores after 30 and 60 days, respectively. 58 participants were diagnosed and selected for the study. Initial quality control showed that the probiotics survived the acidic environment. Over the 60-day period, Oxford Well-Being Questionnaire scores improved (P value < 001).

[0102] Materials and Methods participants All participants provided informed consent for inclusion before participating in the study. Given the nature of the study design as a consumer research study, prior ethics approval was not required. However, the study was conducted in accordance with the principles of the Declaration of Helsinki. Fifty-eight participants were diagnosed and selected for this study. Study participants were anonymized and assigned a unique participant ID.

[0103] Participants completed an eligibility survey, including their name, age, email address, and address. Participants were confirmed to be healthy and free of any serious diagnoses or suspected medical conditions. Inclusion criteria were healthy participants over 18 years of age, willing to participate, and able to provide informed consent. Exclusion criteria included not suffering from any serious health problems, including diagnosed mental health disorders, and not taking any medications, including antibiotic or steroid treatments. Mental health disorders included clinically diagnosed anxiety, depression, and / or stress.

[0104] intervention HN001 was diluted to 9.5 × 10 in the appropriate vehicle. 9 Probiotic capsules containing colony-forming units (CFU) were manufactured. Participants were given sufficient capsules for the 60-day study period. To account for loss of viability over time, participants were required to receive 6 x 10 probiotics per day. 9 ~9.5×10 9 He received CFU.

[0105] Prior to the start of the intervention, a quality check was performed on the probiotic intervention to ensure gastric viability and survival. HN001 capsules were placed in a shaker flask under gastric "mimicking" conditions. Bacteria were shaken at 37°C for 15, 30, and 60 minutes and then tested for viability using standard flow cytometry methods. Briefly, probiotic samples were incubated with permeable (thiazole orange (TO)) and non-permeable (propidium iodide (PI)) DNA-binding dyes to stain all cells and those with compromised membranes, respectively. Compromised bacterial membranes allow PI to enter the cells, while intact, viable cells can exclude them. Therefore, bacterial cells observed to be double-stained with both TO and PI can be considered damaged when their membranes are completely disrupted, allowing more PI into the cells. PI quenches TO, such that dead bacterial cells are only labeled with PI. Using this method, it is possible to distinguish between live, damaged, and dead cells based on the amount of each dye within the cells.

[0106] result The primary outcome was change in Oxford Well-Being Questionnaire (OHQ) scores after 30 and 60 days. The Oxford Well-Being Questionnaire (Argyle et al., 1989) was devised in the late 1980s at the Department of Experimental Psychology at the University of Oxford as a broad measure of personal well-being, primarily for in-house purposes. However, since then, the questionnaire has been widely used, with a review by Argyle, Martin, and Lu in 1995 (Argyle et al., 1995). The scale has been found to function consistently, and other researchers have reported its use in both the UK (Furnham and Brewin, 1990; Joseph and Lewis, 1998) and the US (Valiant, 1993). The OHQ has a total of 29 items. Each item is scored on a Likert scale from 1 to 6 (1 = extremely unhappy, 2 = very unhappy, 3 = somewhat unhappy, 4 = somewhat true, 5 = mostly true, 6 = extremely true). Some items are worded positively and others negatively. Therefore, negative items (1, 3, 12, 13, 16, 18, 21, and 29) should be reverse scored (Hills and Argyle, 2002). The total score is then divided by 29. The interpretation of the final scores is as follows: a final score of 1–2 is considered “unhappy,” a final score of 2–3 is considered “slightly unhappy,” a final score of 3–4 is considered “neutral,” a final score of 4 is “slightly or moderately happy,” 4–5 is “rather happy,” 5–6 is “very happy,” and a final score of 6 is considered “too happy” (Hills and Argyle, 2002).

[0107] Data collection Participants completed the OHQ before receiving the HN001 intervention (Time Point 1), after taking HN001 for 30 days (Time Point 2), and after 60 days (Time Point 3). Participants also answered questions about their age and gender.

[0108] statistical analysis Statistical analyses were performed using SPSS version 26 (Statistics, I., IBM Corp. Released 2013). All statistical tests were two-sided at the 5% significance level. Repeated measures analysis of variance (time effect) was used to evaluate the effect of probiotic use over time. An independent t-test was used to compare the effect of gender on well-being scores.

[0109] result Initial quality inspection and gastric viability Flow cytometry studies performed by BioForm Solutions examined the number of live / damaged / dead cells over three intervals: before acid treatment, 15 minutes after acid treatment, and 30 minutes after acid treatment (Figures 1 and 2). Results showed that HN001 remained viable for 30 minutes after acid exposure at gastric pH. Although the levels of damaged and dead cells increased over time, the decrease in live cells was approximately 0.5 log10, which is considered acceptable. Assuming that the probiotic capsule dissolves quickly in gastric acid and is taken with a small amount of liquid, after 30 minutes, most of its volume (>67%) would have been transported to the small intestine (Goyal et al., 2019).

[0110] Recruitment Volunteers (58) were selected to participate in this study. Each participant was required to complete an eligibility survey to confirm their ability to participate in the study and ensure that the inclusion and exclusion criteria were met. Figure 3 shows the study flowchart. Three participants were lost to follow-up. Eight participants had incomplete data and were excluded from the final analysis. Therefore, the final analysis included 47 participants.

[0111] Baseline data A total of 58 participants were recruited for the study; however, three decided not to participate, and eight participants had incomplete data and were subsequently excluded from the study. Therefore, 47 participants were analyzed. Table 1 shows the demographic characteristics of the participants. The mean age of the participants was 37.7 years, ranging from 21 to 66 years. There were slightly more men than women in the study, but this difference was not statistically significant. The mean (SD) happiness score at baseline was 4.29 (0.65), ranging from 2.97 to 5.55 (Table 2). [Table 1]

[0112] result A summary of the OHQ results at the three time points is shown in Table 2. The results showed a clear increase in the average happiness score over time, with more people shifting beyond the neutral category over time. In fact, 72% of consumers improved their happiness score after 30 days, and 79% improved their happiness score after 60 days compared to baseline. No participants were classified as unhappy at time points 2 or 3. The data were then categorized as unhappy and happy, using "below neutral" as unhappy and "moderately happy" and "rather happy" as happy (Table 3). This classification showed a similar trend of increased happiness. [Table 2] [Table 3]

[0113] An independent t-test showed that there were no significant differences between the mean happiness scores between men and women (Table 4).

[0114] Using repeated measures ANOVA, results showed that there was a statistically significant increase in well-being scores over time. Post-hoc analysis using LSD showed that time points 2 and 3 were statistically different when compared to time point 1, but no significant differences were observed between time points 2 and 3 (Table 5). Figure 4 shows the mean plots and estimated marginal means. [Table 4] [Table 5]

[0115] conclusion The study showed that HN001 survived the initial acidity test, an important factor in any successful probiotic strain (Reid, 2005). The study also demonstrated that participants had an increase in well-being scores over time compared to baseline.

[0116] Example 2 - Effect of probiotics on mental well-being and happiness The primary objective of the study was to determine the effect of HN001 on scores over a 4-week period using the Oxford Well-Being Questionnaire (OHQ). Secondary objectives of the trial included: 1. To determine the effect of HN001 on salivary cortisol levels over a 4-week period. 2. To determine the effect of HN001 on stress, anxiety, and depression scores using the DASS-21 over a 4-week period.

[0117] Materials and Methods Clinical trial design A 30-day double-blind, placebo-controlled, randomized controlled trial.

[0118] Samples and Sampling Participants (120) will be placed in the trial (60 in the probiotic group: 60 in the placebo group). Simple randomization (1:1) will be used. The randomization plan will be maintained by the sponsor and will not be shared with the research team.

[0119] participants The study aims to recruit 120 participants. Inclusion criteria - Adults aged 18-60 -Currently living in the United States -Have access to a smartphone, internet, and computer -Reports moderate levels of stress Participants: -Currently taking medication for depression or anxiety -If you have had antibiotics within the past month -If you are allergic to coconut, you will be excluded.

[0120] Recruitment and eligibility screening All eligibility screening, consent, and data collection will be managed through a secure web-interface data collection platform facilitated by Survey Monkey. Participants will be recruited from a customer database. The advertisement will provide a link and QR code that interested people can use to access the Participant Information Sheet (PIS). Those interested in participating will be asked to complete brief eligibility questions, including a self-report stress questionnaire, the Perceived Stress Scale ("PSS"). The PSS is a 10-item questionnaire asking participants to assess how often they have felt stressed in the past month. Scores range from 0 to 40, with higher scores indicating higher stress levels. Eligible participants will have a score of 14 or greater.

[0121] Eligible participants will be directed to an electronic consent form. After consenting, participants will answer demographic questions, including contact details, which will be provided with the test product. The entire enrollment process will take approximately 10 minutes at a time.

[0122] Baseline data collection Once consent and demographic data collection is complete, participants will be sent a wearable Fitbit Charge 5 device, which they will wear for two weeks according to instructions to provide pre-study reference data. After the two-week Fitbit reference data collection period, participants will receive a link to complete a baseline questionnaire (Survey 1) and a mailed home salivary cortisol test kit (with instructions and a postage-paid return pack). Gut microbiome testing will also be conducted at this time. Participants will receive their study products at this time.

[0123] intervention Participants will be randomly assigned to receive HN001 probiotic capsules or a placebo. Participants will be asked to take the capsules daily for four weeks with a meal or drink. HN001 is the name of the probiotic strain. The placebo is coconut powder. Both probiotic and placebo capsules are dairy-free and gluten-free. Adherence monitoring: Emails or text messages (depending on participant preference) will be sent at weeks 2, 3, and 4 to remind participants to take the supplement daily.

[0124] Midpoint evaluation At the midpoint of the intervention (2 weeks after starting the supplement), participants will be sent a link to a questionnaire (Survey 2).

[0125] Post-intervention evaluation One week before participants were due to complete the intervention, they were sent a saliva collection kit. They were asked to collect a saliva sample and return it using the provided prepaid mailing bag. Another microbiome test was also performed. At the end of the intervention period, participants were sent an electronic link to complete a questionnaire (Survey 3).

[0126] Questionnaire The following questionnaire will be completed by study participants: The Oxford Well-Being Inventory (described above in Example 1) and the Depression, Anxiety, and Stress Scale DASS-21 (Lovibond and Lovibond, 1995).

[0127] Salivary cortisol A saliva kit is included as part of the microbiome collection kit. The saliva kit includes a 15 ml sterile plastic tube with a lid and instructions. The instructions outline that participants should self-collect a saliva sample before bedtime by spitting saliva into the tube, filling it halfway. The collection process should take no more than 15-30 minutes in total. Participants write their name, date, and collection time on the tube. The entire sample volume will be used for measurement.

[0128] Fitbit wearable activity data (from wearable devices): Participants will be asked to wear a Fitbit Charge 5 watch for a total of 16 weeks, consisting of a two-week baseline period and a 14-week intervention period. Participants will be asked to wear it at all times for a total of 14 weeks, except when washing or swimming. Data extraction from the Fitbit will be cleaned and aggregated by a third-party company, DataMine. The raw data will be provided to an independent statistician via encrypted USB stick for analysis.

[0129] Microbiome The microbiome kit includes a 2 ml sterile plastic tube with a lid and instructions. The kit also includes gloves, a fecal collector, and a prepaid USPS envelope. The instructions outline that participants must self-collect the fecal sample. The collection process should not take more than 5 minutes in total. Participants register their sample through the sample portal. The entire sample volume will be used for measurement.

[0130] statistical analysis T-tests will be used to analyze differences between intervention groups in psychological well-being, salivary cortisol, and objective physical activity from Fitbit. Repeated measures analysis of covariates will also be used. Descriptive statistics will be used additionally.

[0131] It will be appreciated by those skilled in the art that numerous variations and / or modifications may be made to the present invention as illustrated in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

[0132] This application claims priority to Australian Provisional Application No. 2022 / 903873 entitled "Methods for increasing mental well-being and / or happiness", filed on December 16, 2022, the entire contents of which are incorporated herein by reference.

[0133] All publications discussed and / or referred to herein are incorporated herein in their entirety.

[0134] Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be assumed that any or all of such matters form part of the prior art or were general knowledge in the art relevant to the present invention existing before the priority date of each claim of this application.

[0135] References Argyle et al (1989) Recent advances in social psychology: An international perspective 189-203. Argyle et al (1995) Stress and emotion 15:173-187. Chatterjee et al (2020) Asian journal of psychiatry 51:102071. Diener et al (1985) Journal of Personality Assessment 49:71-75. FAO / WHO (2001) Evaluation of health and nutritional properties of powder milk and live lactic acid bacteria 1-4. Fordyce (1988) Social indicators Research 20(4):355-381. Furnham and Brewin (1990) Personality and individual differences 11:1093-1096. GBD Mental Disorders Collaborators (2022) Global, regional, and national burden of 12 mental disorders in 204 countries and territories, 1990-2019: a systematic analysis for the Global Burden of Disease Study 2019. The Lancet Psychiatry. Goyal et al (2019) Neurogastroenterology & Motility 31:e13546. Hills and Argyle (2002) Personality and individual differences 33:1073-1082. Joseph and Lewis (1998) Journal of clinical psychology 54:537-544. Lovibond and Lovibond (1995) Manual for the Depression Anxiety & Stress Scales. (2nd Ed.) Sydney: Psychology Foundation. Lyubomirsky and Lepper, (1999). Social Indicators Research 46:137-155. Reid (2005) Current pharmaceutical design 11:11-16. Rossouw et al (2021) PLoS One 16:e0259579. Ryff (1989) Journal of Personality and Social Psychology, 57, 1069-1081. Ryff et al (2007) National Survey of Midlife Development in the United States (MIDUS II), 2004-2006: Documentation of the Psychosocial Constructs and Composite Variables in MIDUS II Project 1. Ann Arbor, MI: Inter-university Consortium for Political and Social Research. Sharma et al (2021) Current microbiology 78:449-463. Sivamaruthi et al (2019) Tropical Journal of Pharmaceutical Research 18:889-895. Slykerman et al (2017) EBioMedicine 24:159-165. Statistics, I., IBM Corp. Released 2013. IBM SPSS Statistics for Windows, Version 26.0. Stewart-Brown et al (2009) Health and quality of life outcomes, 7(1):15. Tay et al (2020) Nutrients 12:3530. Tennant et al (2007) Health and Quality of Life Outcomes, 5(1):63. Valiant (1993) Personality and individual differences 15:447-453. Vinderola et al (2022) Foods 11:1077. Watson et al (1988) Journal of Personality and Social Psychology 54:1063-1070. Zhou et al (2005) International Journal of Feed Microbiology 98:211-217. Zolkiewicz et al (2020) Nutrients 12:2189.

Claims

1. The use of an effective amount of Lacticaseibacillus rhamnosus HN001 or its derivatives or components in the manufacture of a composition for increasing feelings of well-being in a subject.

2. Use according to claim 1, wherein one or more of the following apply: i) Increasing happiness includes treating unhappiness in the subject; ii) Happiness is assessed using one or more of the following: the Oxford Happiness Scale (OHQ), the Subjective Happiness Scale (SHS), the Life Satisfaction Scale (SLS), the Panas Scale (PS), the True Happiness Scale (AHI), the Psychological Well-being Scale (PWS), and the Fordice Sentiment Scale (FEQ); iii) Happiness is assessed using the Oxford Happiness Quantitative (OHQ) mentioned above; iv) Lacticaseibacillus rhamnosus HN001 or its derivatives or components are administered daily; v) The administration period is approximately 17 to 80 days; vi) The administration period is approximately 30 days or approximately 60 days; vii) The subject has never been treated for one or more of the following: anxiety, depression, stress, heightened restlessness, occasional anxiety, postpartum depression, and postpartum anxiety; viiii) The subject is female; ix) The subject is male; x) The subject is not pregnant, prenatal, or postpartum; xi) The subject has never been pregnant; xi) The subject is a human being; xiiii) The subject is a companion animal or a domestic animal; xiv) The subject is selected from infants, children, adolescents, adults, and the elderly; xv) Lacticaseibacillus rhamnosus HN001 or its derivatives or components are in a form selected from tablets, powders, capsules, liquids, syrups, gummies, and sachets; xvi) Lacticaseibacillus rhamnosus HN001 or its derivatives or components are administered in the form of a nutritional supplement or food; and xvii) Lacticaseibacillus rhamnosus HN001 or its derivatives or components are viable at the time of administration.

3. The use according to claim 1, wherein increasing well-being in the subject includes changes in salivary cortisol levels.

4. The use according to claim 1, wherein increasing well-being in the subject does not involve changes in salivary cortisol levels.

5. The use of the method according to claim 1, comprising one or more of the following: i) Approximately 6 x 10 per day ９ ~Approx. 1×10 １０ This includes administering the colony-forming unit (CFU) Lacticaseibacillus rhamnosus HN001 or a derivative thereof; ii) Approximately 6 x 10 per day ９ Administer the colony-forming unit (CFU) Lacticaseibacillus rhamnosus HN001 or a derivative thereof; iii) The administration is by oral administration; iv) further comprising administering one or more other probiotics or postbiotic microorganisms.

6. The one or more other probiotics or postbiotic microorganisms mentioned above i) Acetilactobacillus, Agrilactobacillus, Akkermansia, Amylolactobacillus, Anaerobut yricum, Apilactobacillus, Bacillus, Bacteroides, Bombilactobacillus, Christensenella, Clostridium, Companilactobacillus, Dellaglioa, Enterococcus, Eubacterium, Faecalibact erium, Fructilactobacillus, Furfurilactobacillus, Holzapfelia, Lacticaseibacillus, La ctiplantibacillus, Lactococcus, Lapidilactobacillus, Latilactobacillus, Lentilactob acillus, Leuconostoc, Levilactobacillus, Ligilactobacillus, Limosilactobacillus, Liqu orilactobacillus, Loigolactobacillus, Parabacteroides, Paucilactobacillus, Pediococcus selected from S, Prevotella, Schleiferilactobacillus, Secundilactobacillus, and Streptococcus; ii) Akkermansia muciniphila, Anaerobutyricum hallii, Bacillus coagulans, Bacillus subtilis, Bacteroides fragilis, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Christensenella minuta, Clostridium butyricum, Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Faecalibacterium prausnitzii, Lactic casei bacillus, Lactic casei bacillus Paracasei, Lacticaseibacillus rhamnosus, Lactiplantibacillus pentosus, Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactococcus lactis, Latylactobacillus sakei, Lentilactobacillus kefiri, Leuconostoc mesenteroids, Levilactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, Limosilactobacillus reuteri、Parabacteroidesselected from Goldsteinii, Pediococcus acidilactic, Pediococcus pentosaceus, Prevotella copri, Saccharomyces boulardii, and Streptococcus thermophilus; iii) L. reuteri RC-14, B. longum BB536, L. lactis MG1363, L. plantarum LP299V, L. plantarum LP-115, B. lactis HN019, L. reuteri JCM1112, L. casei ATCC393, B. animalis B94, L. paracasei LPC-37, L. acidophilus NCFM, B. lactis BL-04, B. lactis Bi-07, L. crispatus LBV88, L. Selected from gasseri LG-36, B. longum CCFM1029, L. rhamnosus HN067, L. rhamnosus GG (LGG), B. brevere M-16V, B. lactis BB12, and L. acidophilus HN017; and / or iv) The use according to claim 5, wherein the psychobiotic is used.

7. The use according to claim 1, further comprising administering one or more prebiotics.

8. The use according to claim 1, wherein the prebiotic is selected from one or more of galacto-oligosaccharides, fructo-oligosaccharides, trans-galacto-oligosaccharides, inulin, human milk oligosaccharides, gentio-oligosaccharides, xylo-oligosaccharides, isomalt-oligosaccharides, bran, resistant starch, lactulose, lactitol, flavanol, and pectin oligosaccharides.

9. Use according to claim 2, wherein one or more of the following apply: i) The food is a dairy product; ii) The dairy product is selected from milk, yogurt, drinking yogurt, butter, cream, ice cream, and cheese; and iii) The food or nutritional supplement is selected from infant formula, follow-up formula, growth formula, breast milk formula, breast milk supplement, adult nutritional formula, and elderly nutritional formula.

10. Use of Lacticaseibacillus rhamnosus or its components, including the AscI restriction enzyme digestion profile of Lacticaseibacillus rhamnosus HN001 shown in Figure 5, in the manufacture of a composition for increasing well-being in a subject.

11. Use of Lacticaseibacillus rhamnosus HN001 or its derivatives or components in the manufacture of compositions for increasing mental well-being in subjects.

12. Use of Lacticaseibacillus rhamnosus or its components, including the AscI restriction enzyme digestion profile of Lacticaseibacillus rhamnosus HN001 shown in Figure 5, in the manufacture of a drug for increasing mental well-being in a subject.

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